A novel heterozygote allele in caprine melanocortin 1 receptor (MC1R) gene: an association with heat stress traits.

Climate change negatively influences the productive and reproductive abilities of goats. There is a need to understand the relationship between heat stress and genes that may aid in the development of climate-resilient goats. Melanism variation in goats plays a role in thermoregulation, in which the melanogenic genes have a pleiotropic effect on the regulation of physiological responses and behavior that are altered due to heat stress in the animals. Thus, the present study was conducted to establish a possible association between the coat color gene (MC1R) and heat stress characteristics. The physiological responses and cortisol levels were recorded in forty different coat-colored goats. The genotyping of the animals revealed four SNPs at the 183rd (C/T), 332nd (C/G), 748th (G/T), and 801st (C/G) positions, among which the black and brown goat populations had novel SNPs at the 332nd position. Eight haplotypes were constructed, and an association study revealed that haplotypes (CCGG, TCGG, and CCTC) that were linked to white animals had lower cortisol values, rectal temperature, skin temperature, and respiration rate. The multivariate and cluster analyses revealed that the white goats were distinct from the rest of the goats. In addition, the docking results revealed the residues that were forming the interaction complex, which could play a role in melanogenesis in the animals and, in turn, the heat stress ability of the goats. Altogether, the results of the present study could pave the way for more research into coat color genes and their relationship with heat stress traits.

Molecular characterization and prevalence of bovine hemoprotozoan and rickettsial organism from Bhubaneswar, Eastern India.

The blood parasites adversely affect the bovine population and are one of the major hindrances to productivity and health of livestock, globally. This study aimed to study the prevalence of some bovine hemoparasites, their molecular characterization, and associated risk factors in and around Bhubaneswar, a place in Eastern India. Microscopy (Giemsa stained thin blood smear) and polymerase chain reaction (PCR)-based prevalence studies were undertaken for a total of 106bovine blood samples belonging to different breed, age, and sex presented to Veterinary Clinical Complex. DNA was extracted by conventional method and primer targeting msp5 gene (382 bp) for Anaplasma marginale, Tams1 gene (572 bp) for Theileria annulata and VSG RoTat 1.2 gene (110 bp) for Trypanosoma evansi, were used. Sequencing and phylogenetic analysis were also carried out for representative PCR products. The risk factors were correlated with prevalence by logistic regression analysis. Blood smear examination revealed a prevalence an overall of 48.11% (51/106) out of which the prevalence of T. annulata, A. marginale and T. evansi, and concurrent infection were reported to be 26.42%, 10.38%, 3.77%, and 7.55% respectively. All the samples found positive in microscopy were also found positive in PCR. PCR assay revealed an overall prevalence of 73.58% (T. annulata: 37.74%, A. marginale:17.92%, T. evansi: 5.66% and concurrent infection:12.26%). A higher prevalence was observed in the age group of 2-4 years (37.74%), females (61.32%) and cross-bred Jersey (50%). Representative PCR products were sequenced and assigned by GenBank (OL550058: A. marginale; OL604429: T. annulata and OL550059: T. evansi). There is a paucity of disease mapping of these parasites in most parts of the country. The study is probably the first report of molecular characterization of T. evansi and A. marginale from the region which gives some information of possible genetic diversity of the isolates circulating in the region. Future research should include large-scale epidemiological surveys as well as concurrent diagnosis of these hemoparasites for better treatment and control strategies.

Molecular epidemiology of lumpy skin disease outbreak in Odisha, India.

Lumpy skin disease virus (LSDV) is the causative agent of lumpy skin disease (LSD) which is a member of Capripoxvirus. It is an economically critical transboundary disease affecting cattle. This study records an LSD outbreak in Ganjam district of Odisha, India during August 2020. The epidemiological data were analysed and LSDV was genetically characterized. Out of the 452 animals clinically examined (59 farms), 63 animals were clinically affected with LSD, with a total morbidity rate of 13.93%. The morbidity rates in the surveyed villages (n = 10) varied from 5.55 to 21.62%. The multivariable logistic regression analysis showed that grazing of animals (P = 0.013; OR: 2.04; 95% CI: 1.16-3.57) and age of cows > 3 years old (P = 0.001; OR: 2.90; 95% CI: 1.65- 5.07) were potential risk factors for the presence of LSD. Out of the 53 clinically suspected animals' samples, 18 samples (33.96%) were found positive for both the P32 and F genes of Capripoxvirus by PCR. Phylogenetic analysis of the P32 gene of LSDV (MW147486) showed 100% similarity with other isolates from India, Bangladesh, Egypt and Saudi Arabia. Additionally, phylogenetic analysis of the F gene of LSDV (MW147485) revealed a similarity of 97.99%, with Odisha India (MT074110) isolate and located in the same cluster with other Indian isolates.

Identification and futuristic role of novel polymorphism of caprine PrP gene.

The Caprine Prion Protein (PrP) gene polymorphism in three different native Indian goat populations of Southern Odisha, namely Ganjam (a registered breed of India), Ghumusari and Raighar was studied. The 876 bp amplified segment of PrP gene contains full length coding sequence of 771 bp. In Ganjam and Ghumusari goats, any difference of nucleotide sequence was not identified. However, the comparison of nucleotide sequences of Raighar goats and goats of other locality revealed a change in nucleotide at five different positions (G190A, G724A, A727T, C775G and C800T) which includes two non-synonymous nucleotide changes. The non-synonymous nucleotide change resulted a change in amino acid at two different positions (Ser234Cys and Lys246Phe) in mature polypeptide which were not reported earlier and therefore, considered as novel. On the basis of these variants of PrP gene phylogenetic tree was constructed which showed that Ganjam and Raighar goats appeared in different clade. Since any occurrence of Scrapie infection in goats of Odisha was not reported, it can be proposed that these changes in amino acid may be responsible as resistance allele.

Transplacental transmission of Theileria annulata in cattle confirmed by molecular techniques.

Theileria annulata (T. annulata) is a tick-borne apicomplexan parasite affecting ruminants and causes severe economic loss. The present paper reports the vertical transmission of T. annulata in a three day old crossbred Jersey calf. Molecular diagnosis of tropical theileriosis was done by PCR, cloning, sequencing and phylogenetic analysis of isolated parasites. Calf showed very low hematocrit value (HCT) (17.82%), red blood cells count (RBC) count (6.9 × 106/µL), and hemoglobin concentration (Hb) (6.6 g/dL). Phylogenetic analysis of Tams 1 gene showed that T. annulata Khorda isolate (MW123091) shared 99.23%, 99.23% and 99.11% nucleotide homology with Puri, India (MN818858), Bahrain (AF214797) and Hyderabad, India (MK034702), respectively. This is the first study in Odisha, India to give an insight into the molecular detection, phylogeny and hematological analysis of T. annulata infected crossbred Jersey calf which got the infection through transplacental transmission.

Molecular detection of Theileria annulata infection in cattle by conventional PCR and quantitative real time PCR in India.

Theileria annulata (T. annulata) is a tick-borne apicomplexan parasite that affects bovine. It is endemic in many tropical and subtropics areas, including Odisha, India. The objective of this study is to identify T. annulata infection in the peripheral blood of cattle as a biological sample by conventional PCR (cPCR) and quantitative PCR (qPCR). The phylogenetic analysis was done using the T. annulata merozoite surface antigen (Tams 1) gene. Out of 552 samples of examined blood smears by microscopy, 454 (82.24%) animals were positive for Theileria species. Out of 454 samples, 96 samples were further examined by both cPCR and qPCR, 52 samples (54.16%) were found positive for T. annulata in both PCR methodologies. Phylogenetic analysis revealed that T. annulata Odisha isolate was closely related to T. annulata Uttarakhand, India isolate (KM061799) and Hyderabad, India isolate (MK034702) with Nucleotide sequence identity 95.36%, 95.25%, respectively. This is the first study to detect T. annulata by qPCR in Odisha and supported that both PCR techniques were equally effective for the detection of Tams 1 gene of T. annulata in cattle's blood.

Novel polymorphism of HSP70 gene affected caprine physiological vital parameters.

The synthesis of heat shock protein 70 (HSP70) is temperature-dependent and its response is considered as a cellular thermometer in response to thermal stress. The variation in HSP70 gene expression has been positively correlated with thermo-tolerance. Three different goat populations (Black Bengal, Ganjam and Raighar) which differ in body size, coat color and production performance were assessed for effect of thermal stress at different temperature humidity index (THI). The physiological vital parameters like rectal temperature (RT), skin temperature (ST), heart rate (HR) and respiration rate (RR) which are related to thermal stress susceptibility were recorded. The genetic variation of HSP70 gene in the three goat populations and its effect on physiological vital parameters related to thermal stress was assessed. A novel polymorphism (C241T) in the exonic region of HSP70 gene with significant (p < .05) statistical association with the physiological vital parameters like RT, ST and RR was identified indicating its role in thermo-tolerance.HighlightsThe HSP70 gene was found to be polymorphic in Indian goats.The non-synonymous nucleotide change (C241T) resulted change in amino acid Arginine 241 Cysteine in mature polypeptide which were not reported earlier.The constructed phylogenetic tree showed that Ganjam and Raighar goats are more close to each other.

Molecular, epidemiological and haematological evaluation in Theileria orientalis infected cattle from an endemic region in India.

The objective of the present study was to investigate the epidemiological and haematological parameters with simultaneous molecular detection of Theileria orientalis infection of crossbred jersey (CBJ) cattle. Haematological values like mean hemoglobin (Hb) (7.31 ± 2.3 g/dl), packed cell volume (PCV) (21.69 ± 6.11%), red blood cells count (RBCs) (4.40 ± 1.6 M/µl), white blood cells count (WBCs) (6.93 ± 3.06 103/µl) and mean corpuscular hemoglobin concentration (MCHC) (33.56 ± 3.51 g/dl) were decreased significantly (p < 0.05), whereas mean corpuscular volume (MCV) (51.06 ± 6.14fl) and eosinophil count (0.39 ± 0.44 103/µl) were significantly (p < 0.05) increased in cattle infected with T. orientalis. Analysis of major piroplasm surface protein (MPSP) of 110 blood samples randomly collected from cattle from seven districts by PCR indicated that an average of 70% of cattle was positive for T. orientalis infection. In particular, Puri and Khorda districts were identified as relatively high-risk areas for T. orientalis infection, with infection rates of 76.66% and 72.4%, respectively. The phylogenetic analysis of isolated T. orientalis MPSP gene (MN334767) classified it into type 5. Earlier Indian isolates were classified into three types viz.type 1, type 3 and type 7 and this is the first time to detect type 5 in Odisha, India.

An integrative bioinformatics analysis of caprine GNAI3 gene.

Goat is the most preferred domesticated animal in Indian subcontinent. However, the climatic change-induced heat stresscauses a formidable challenge for maintaining optimum productivity. G protein subunit alpha i3 (GNAI3) is one of the genes that may have significant role in heat tolerance mechanism in goats. The caprine GNAI3 gene was searched for homology analysis and its three dimensional protein structure was predicted followed by its validation through in silico approach. Nucleotide sequence-based phylogenetic tree analysis showed that the caprine GNAI3 gene has close evolutionary relationship with that of Ovis aries. Homology modelling of caprine GNAI3 protein was done in MODELLER 9.18 (P1), PHYRE2 (P2), GENO3D (P3) and SWISS MODEL (P4). The modelled structures were further validated after observing the Ramachandran and hydrophobicity plots. In the best of three dimensional protein structure (P4 as produced by SWISS MODEL), 330 (98.8%), three (0.9%) and one (0.3%) amino acid residues were found in favoured region, allowed region and outlier region, respectively. Degree of hydrophobicity of the generated protein structures revealed the presence of alternate hydrophobic and hydrophilic regions. The ligand receptor interaction site of the predicted 3D model was traced out using Discovery Studio 3.5. STRING database revealed protein interactions with Plcb1, Plcb2, Plcb3 and other proteins of G family such as Gnb1, Gnb2, Gnb3,Gnb4, Gng2, Gng4 and Gpsm1. KEGG pathway maps revealed interaction with eNOS, iNOS, VEGF and MAPK, which are reported to be transcribed in response to heat stress. Thus, caprine GNAI3 can be used as a possible biomarker for studying heattolerance mechanism in goats.

An in vivo and in silico analysis of novel variation in TMBIM6 gene affecting cardiopulmonary traits of Indian goats.

Transmembrane Bax Inhibitor Motif-containing 6 (TMBIM6) gene acts as calcium leak channel and negatively regulates autophagy and autophagosome formation. The TMBIM6 gene was amplified and searched for variation in three different goat populations (i.e. Black Bengal, Ganjam and Raighar) of Odisha state of the India. The result indicated two substitutions i.e. 55th position (C55T) and 95th position (C95A) in the amplified region of the gene resulting in change of amino acids (Leu > Phe and Thr > Asn). The identified SNPs were combined to form haplotypes and animals were grouped accordingly. Structural analysis showed minor changes (5%) in between mutant and wild TMBIM6 protein structures. However, any functional variation could not be identified with respect to the calcium ligand and open pore state. But an alteration of calcium binding site was found. The binding interaction of calcium with the TMBIM6 protein was hydrophobic in nature in closed state whereas hydrophilic in open pore stage. The stress releasing function was the result of calcium leakage controlled by amino acids coded by exon 4 and exon 5 regions of TMBIM6 gene. The effect of breed and haplotype on cardiopulmonary traits was studied. The data on cardiopulmonary traits of body i.e. rectal temperature, skin temperature, heart rate and respiration rate were recorded when ambient temperature usually remained the highest. The statistical analysis showed, significant difference in rectal temperature, skin temperature and respiration rate among these goat populations. The haplotypes (CC and TA) were found to have a significant (P < 0.05) effect on rectal temperature, skin temperature and respiration rate. However, any such significant effect could not be identified in recorded heart rate. The objective of the present study to identify the genetic variations in TMBIM6 gene having significant effect on cardiopulmonary traits which can be further uses as the molecular markers to improve heat tolerance mechanism in goats.

In silico analysis of caprine superoxide dismutase 1 (SOD1) gene.

Climatic change induced heat stress causes a formidable challenge for maintaining optimum productivity in goats. The SOD1 gene (superoxide dismutase 1) is one of the genes that regulates the heat tolerance capacity. This study aimed to in silico characterize this gene. Sequence based phylogenetic tree analysis showed the caprine SOD1 gene has close evolutional relationship with that of sheep. STRING database reveals its interaction with SOD2 (Superoxide dismutase [Mn], mitochondrial), ATOX1 (Copper transport protein), RAC1 (Ras-related C3 botulinum toxin substrate 1), GPX2 (Glutathione peroxidase 2), GPX4 (Glutathione peroxidase 4), GPX5, GPX6, CAT (Catalase). The KEGG pathway maps gave interaction with eNOS, iNOS, VEGF and MAPK in which gene transcription modulates response to heat stress. The three dimensional protein structure was predicted using Modeller, Phyre 2, and Swiss Model. Structure validation was done observing the Ramachandran Plot and Hydrophobicity Plot in Discovery Studio considering amino acid residues in favoured region.

Meta-analysis of seminal traits affecting bull fertility.

The pedigree information on bulls was collected from Frozen Semen Bank (FSB). Male reproductive data on seminal characters (semen volume (SV), sperm concentration (SC), initial progressive motility (IPM), post-thaw motility (PTM), and total spermatozoa (TS)) were analyzed to calculate their repeatability, heritability, genetic correlation, phenotypic correlation, and environmental correlation values. The principal component analysis revealed SV and SC were the two most important characters among the five seminal characters that can be further used. The repeatability value for seminal characters was very low (0.02-0.2). The sire component of heritability value varied from very low to low (0.0001 to - 0.24). The dam component of heritability value varied from very low to high (0.01 to 0.64). The genetic correlation value calculated from the sire component indicated a slightly antagonistic relationship between SV and SC (- 0.007) and SV and PTM (- 0.049). The environmental correlation value was found to be negative for all the traits except the value calculated from the dam component between SC and IPM (0.027). The phenotypic correlation values between the seminal traits were low and mostly negative except between SC and IPM (0.107) and PTM and TS (0.109). Statistically significant correlation was observed between PTM and SV (- 0.157), PTM and IPM (- 0.145), PTM and TS (0.109), and SC and IPM (0.107). Since the estimated heritability, repeatability, and the correlation values were found low, individual selection method cannot be applied on these bulls.

Molecular Characterization of Mx1 Gene in Native Indian Breeds of Chicken.

The genetic polymorphism of Mx1 gene was explored in Indian chicken breeds. PCR-RFLP analysis in 102 bp fragment of partial intron 13 and partial exon 14 of Mx1 gene revealed two genotypes viz. RS and SS with two alleles viz. R and S both in Naked Neck and Tellicherry breeds of chicken. The homozygous genotype RR was not identified. When deduced amino acid sequences were compared, the asparagine amino acid was found to be substituted in "R" allele for serine in "S" allele. PCR-SSCP analysis of 284 bp fragment in 5'-UTR and partial promoter region revealed three genotypes viz. CC, CG, and CH with three different alleles viz. C, G, and H in Naked Neck breed of chicken and five genotypes viz. DI, JK, KK, KL, and KM with six different alleles viz. D, I, J, K, L, and M in Tellicherry breed of chicken. The homozygous genotypes viz. GG and HH in Naked Neck and DD, II, JJ, LL, and MM in Tellicherry chicken was not identified. The nucleotide substitution rate estimated to be in the range of 0.004-0.011. The identified genetic variation can be helpful for better insight to disease resistance property of the Mx1 gene.

Identification of sperm motility markers in bovine transition protein genes.

Transition proteins (TNPs) are essential in chromatin condensation during spermiogenesis, and hence, they are the candidate genes for identifying sperm motility markers. Coding and in silico predicted promoter regions of these genes were investigated in crossbred and purebred cattle, and also, their mRNA quantification was done to explore its use as a diagnostic tool of infertility. PCR-SSCP analysis revealed two band patterns in fragment III of TNP1 and fragment II of TNP2 gene. Sequence analysis revealed a deletion of "G" nucleotide in 3'UTR region of TNP1 and C>T SNP in intronic region of TNP2 gene. Least square analysis of variance did not reveal any significant influence of nucleotide deletion on any sperm motility parameters in both crossbred and purebred cattle. However, C>T SNP had a significant effect on initial progressive motility (p < 0.05) in purebred cattle and post-thaw motility in overall cattle population. RT-qPCR analysis did not reveal any significant variation in TNP1 and TNP2 gene expression among poorly motile and good quality spermatozoa of Vrindavani bulls.

Nucleotide variability of protamine genes influencing bull sperm motility variables.

Protamines (PRMs), important proteins of chromatin condensation in spermiogenesis, are promising candidate genes to explore markers of sperm motility. The coding and in-silico predicted promoter regions of these genes were investigated in 102 crossbred and 32 purebred cattle. Also, mRNA quantification was done to explore its possibility as diagnostic tool of infertility. The PCR-SSCP analysis indicated there were two band patterns only in fragment I of the PRM1 and fragment II of the PRM2 gene. The sequence analysis revealed A152G and G179A transitions in the PRM1 gene. Similarly, G35A, A49G and A64G transitions were identified in the PRM2 gene which resulted in altered amino acid sequences from arginine (R) to glutamine (Q), from arginine (R) to glycine (G) and from arginine (R) to glycine (G), respectively. This caused the reduction in molecular weight of PRM2 from 2157.66 to 1931.33 Da due to reduction in the number of basic amino acids. These altered properties of the PRM2 protein led to the reduction in Mass Motility (MM: P < 0.01), Initial Progressive Motility (IPM; P < 0.05) and Post Thaw Motility (PTM; P < 0.05) in crossbred bulls. The least squares analysis of variance indicated there was an effect of PRM2 haplotypes on MM (P = 0.0069), IPM (P = 0.0306) and PTM (P = 0.0500) in crossbred cattle and on PTM (P = 0.0408) in the overall cattle population. Based on the RT-qPCR analysis, however, there was not any significant variation of PRM1 and PRM2 gene expression among sperm of Vrindavani bulls with relatively lesser and greater sperm motility.

Expression Profiling and Identification of Novel SNPs in CatSper2 Gene and Their Influence on Sperm Motility Parameters in Bovines.

122 randomly selected Vrindavani cattle were studied to detect polymorphism in four fragments of the CatSper2 gene that were comprised of exon 2, 4, 5, and 6 with flanking regions. Using PCR-SSCP and sequencing analysis, three SNPs (T157C, C273A, and A274C) in the first fragment, one SNP (C30G) in the second fragment, and two SNPs (T86G and T292C) in the fourth fragment were identified. The third fragment did not reveal any polymorphism. The SNPs were used for construction of haplotypes and three haplotypes were found. The least square analysis of variance revealed a significant (P < 0.01) effect of haplotype on all three motility parameters. The haplotype II and III were nonsignificantly different from each other while being significantly (P < 0.01) different from haplotype I. The nonsignificant difference of haplotype II with III can lead to a hypothesis that T>G or C>T SNPs may not play a role in sperm motility. However, when the comparison was made between haplotype I and II, it can be inferred that C>T SNP may have a role in sperm motility, as haplotype II has better motility parameters. Expression profiling of Catper2 gene revealed nonsignificant down regulation of CatSper2 gene in poor motility sperm compared to good motility sperm.

Single Nucleotide Polymorphisms in 5' Upstream Region of Bovine TLR4 Gene Affecting Expression Profile and Transcription Factor Binding Sites.

The present study in the 5' upstream region of TLR4 gene revealed four Single Nucleotide Polymorphisms (SNPs) in Vrindavani and Tharparkar cattle. The polymorphic information content (PIC), heterozygosity and allelic diversity values were low to moderate for these SNPs. In Vrindavani cattle, one SNP was found to be in Hardy-Weinberg Equilibrium (HWE) and the remaining three were found to be in linkage disequilibrium (LD) as indicated statistically (P > 0.05). In Tharparkar cattle, two SNPs were found to be in HWE and were not in LD as indicated statistically (P > 0.05). These SNPs were used for construction of haplotypes. In-silico analysis of these SNPs predicted abolition of eight transcription factor binding sites and creation of eight new sites. The quantitative real time PCR analysis did not show any significant variation of gene expression among haplotypes. However, gene expression between breed was found to be significant (P < 0.05) which suggested that upstream region of bovine TLR4 gene has a crucial role in its expression. These findings in TLR4 gene offer essential evidence that can be useful in future research exploring its role in immunity. TLR4 can be used as a marker for selection for disease resistance in bovines.

Molecular Cloning, Expression and Characterization of Para Flagellar Rod Protein 1 of Trypanosoma evansi.

BACKGROUND: Antigenic variation allows the trypanosomes to evade the potentially destructive host immune response and is an important reason for failure to develop a protective vaccine. Among the non-variant structural proteins, paraflagellar rod protein (PFR) is a prospective vaccine target owing to its role in the active movement of the parasite. METHODS: The PFR1 gene was cloned in pET-32a expression vector and after confirmation by restriction digestion, expressed as a Histidine-tagged fusion protein, in BL21 DE3 strain of E. coli. The expressed protein was affinity purified and then renatured. The immunoreactivity of the expressed recombinant protein was shown by western blot analysis using the specific serum. The experiment was carried out during 2013-14 at Division of Parasitology, Indian Veterinary Research Institute, Izatnagar, U.P., India. RESULTS: The results of sequencing, restriction digestion analysis, and PCR reaction revealed that cloning of PFR1 gene in pET-32a expression vector and the results of SDS PAGE and Western blot further confirmed its homogeneity and purity. The in silico Te-PFR1 (T. evansi PFR1) nucleotides sequence analysis revealed its close homology with the other members of the order Kinetoplastida. CONCLUSION: We report here the molecular cloning, heterologous expression, and characterization of PFR1, a constituent protein of PFR. Due to its conserved nature, the PFR1 protein could be a prospective vaccine target against multiple Trypanosoma species.

Sequence Based Structural Characterization and Genetic Diversity Analysis of Full Length TLR4 CDS in Crossbred and Indigenous Cattle.

The exploration of candidate genes for immune response in cattle may be vital for improving our understanding regarding the species specific response to pathogens. Toll-like receptor 4 (TLR4) is mostly involved in protection against the deleterious effects of Gram negative pathogens. Approximately 2.6 kb long cDNA sequence of TLR4 gene covering the entire coding region was characterized in two Indian milk cattle (Vrindavani and Tharparkar). The phylogenetic analysis confirmed that the bovine TLR4 was apparently evolved from an ancestral form that predated the appearance of vertebrates, and it is grouped with buffalo, yak, and mithun TLR4s. Sequence analysis revealed a 2526-nucleotide long open reading frame (ORF) encoding 841 amino acids, similar to other cattle breeds. The calculated molecular weight of the translated ORF was 96144 and 96040.9 Da; the isoelectric point was 6.35 and 6.42 in Vrindavani and Tharparkar cattle, respectively. The Simple Modular Architecture Research Tool (SMART) analysis identified 14 leucine rich repeats (LRR) motifs in bovine TLR4 protein. The deduced TLR4 amino acid sequence of Tharparkar had 4 different substitutions as compared to Bos taurus, Sahiwal, and Vrindavani. The signal peptide cleavage site predicted to lie between 16th and 17th amino acid of mature peptide. The transmebrane helix was identified between 635-657 amino acids in the mature peptide.

Single nucleotide polymorphism mining and nucleotide sequence analysis of Mx1 gene in exonic regions of Japanese quail.

AIM: An attempt has been made to study the Myxovirus resistant (Mx1) gene polymorphism in Japanese quail. MATERIALS AND METHODS: In the present, investigation four fragments viz. Fragment I of 185 bp (Exon 3 region), Fragment II of 148 bp (Exon 5 region), Fragment III of 161 bp (Exon 7 region), and Fragment IV of 176 bp (Exon 13 region) of Mx1 gene were amplified and screened for polymorphism by polymerase chain reaction-single-strand conformation polymorphism technique in 170 Japanese quail birds. RESULTS: Out of the four fragments, one fragment (Fragment II) was found to be polymorphic. Remaining three fragments (Fragment I, III, and IV) were found to be monomorphic which was confirmed by custom sequencing. Overall nucleotide sequence analysis of Mx1 gene of Japanese quail showed 100% homology with common quail and more than 80% homology with reported sequence of chicken breeds. CONCLUSION: The Mx1 gene is mostly conserved in Japanese quail. There is an urgent need of comprehensive analysis of other regions of Mx1 gene along with its possible association with the traits of economic importance in Japanese quail.